Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA
Department of Veterinary Microbiology and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames, Iowa, USA.
Infect Immun. 2020 Jun 22;88(7). doi: 10.1128/IAI.00245-20.
Small noncoding RNAs (ncRNAs) are involved in many important physiological functions in pathogenic microorganisms. Previous studies have identified the presence of noncoding RNAs in the major zoonotic pathogen ; however, few have been functionally characterized to date. CjNC110 is a conserved ncRNA in , located downstream of the gene, which is responsible for the production of the quorum sensing molecule autoinducer-2 (AI-2). In this study, we utilized strand specific high-throughput RNAseq to identify potential targets or interactive partners of CjNC110 in a sheep abortion clone of These data were then utilized to focus further phenotypic evaluation of the role of CjNC110 in motility, autoagglutination, quorum sensing, hydrogen peroxide sensitivity, and chicken colonization in Inactivation of the CjNC110 ncRNA led to a statistically significant decrease in autoagglutination ability as well as increased motility and hydrogen peroxide sensitivity compared to the wild-type. Extracellular AI-2 detection was decreased in ΔCjNC110; however, intracellular AI-2 accumulation was significantly increased, suggesting a key role of CjNC110 in modulating the transport of AI-2. Notably, ΔCjNC110 also showed a decreased ability to colonize chickens. Complementation of CjNC110 restored all phenotypic changes back to wild-type levels. The collective results of the phenotypic and transcriptomic changes observed in our data provide valuable insights into the pathobiology of sheep abortion clone and strongly suggest that CjNC110 plays an important role in the regulation of energy taxis, flagellar glycosylation, cellular communication via quorum sensing, oxidative stress tolerance, and chicken colonization in this important zoonotic pathogen.
小非编码 RNA(ncRNAs)参与了病原微生物的许多重要生理功能。先前的研究已经在主要的人畜共患病病原体中发现了非编码 RNA 的存在,然而,迄今为止,仅有少数得到了功能表征。CjNC110 是 中的保守 ncRNA,位于 基因的下游,负责产生群体感应分子自诱导物-2(AI-2)。在这项研究中,我们利用链特异性高通量 RNAseq 鉴定了 CjNC110 在绵羊流产克隆 中的潜在靶标或交互伙伴。然后,利用这些数据进一步对 CjNC110 在运动性、自聚集、群体感应、过氧化氢敏感性和鸡定植中的作用进行表型评估。CjNC110 ncRNA 的失活导致自聚集能力显著下降,与野生型相比,运动性和过氧化氢敏感性增加。ΔCjNC110 中检测到的细胞外 AI-2 减少;然而,细胞内 AI-2 积累显著增加,表明 CjNC110 在调节 AI-2 的转运中起着关键作用。值得注意的是,ΔCjNC110 也表现出降低的定植鸡的能力。CjNC110 的互补恢复了所有表型变化回到野生型水平。我们的数据中观察到的表型和转录组变化的综合结果为 绵羊流产克隆的病理学提供了有价值的见解,并强烈表明 CjNC110 在调节趋化运动、鞭毛糖基化、通过群体感应进行细胞通讯、耐氧化应激和在这种重要的人畜共患病病原体中鸡定植方面发挥重要作用。